SEQanswers

Go Back   SEQanswers > General



Similar Threads
Thread Thread Starter Forum Replies Last Post
Doing PCR enrichment on PCR-free TruSeq lib? JBKri Sample Prep / Library Generation 2 10-08-2015 08:00 AM
PCR based test for TruSeq library contamination in Pre-PCR lab? lescagasan Core Facilities 2 12-21-2013 10:06 AM
PCR-Free Libraries & Bioinformatic PCR Duplicate detection? aeonsim Bioinformatics 3 10-23-2013 02:36 PM
Lib prep - Splitting PCR to reduce PCR bias prepagam Illumina/Solexa 2 03-15-2013 04:30 PM
Number of PCR How many PCR cycles to enrich adapter-modified DNA fragments MGH Man Sample Prep / Library Generation 5 07-26-2010 06:15 AM

Reply
 
Thread Tools
Old 08-14-2017, 04:26 AM   #1
julyduva
Junior Member
 
Location: Colombia

Join Date: Aug 2017
Posts: 1
Default Help with PCR

Good morning,

I'm student of master and my thesis is with animals virus. I have problems with the PCR: I ruled out contamination of the reactives, but in the positive samples appear a unknown band, the negative control is negative.

Can somebody help me with papers or handbooks that talk about mistakes in PCR?

Thanks for all.
julyduva is offline   Reply With Quote
Old 08-21-2017, 05:25 AM   #2
asorbie
Junior Member
 
Location: Munich

Join Date: Jun 2017
Posts: 2
Default

Can you give us a little more information, it may be that the unknown band is due to non-specific binding of your primers? What size is it roughly? What cycling conditions did you use, composition of your master mix etc? Bio-rad PCR troubleshooting is always a good place to start: http://www.bio-rad.com/en-uk/applica...roubleshooting
asorbie is offline   Reply With Quote
Old 08-21-2017, 11:39 PM   #3
Dorota
Junior Member
 
Location: The Netherlands

Join Date: May 2017
Posts: 6
Default

In your positive samples, do you see your expected band AND an unknown band or you just see unknown/unexpected band? Is it PCR that is known to produce good results or you are trying to develop a PCR ?
Dorota is offline   Reply With Quote
Reply

Tags
pcr artifacts, pcr errors

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off




All times are GMT -8. The time now is 06:58 AM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2017, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO